Ad.DCs and mDCs express tmTNF and trans-IL-15. (A) tmTNF and trans-IL-15 expression by iDCs, Ad.DCs (500 MOI), and mDCs was analyzed by imaging flow cytometry. Images show examples of individual cells, and the MFI values listed are averaged for the population of 10 000 cells. (B) Topography of tmTNF and trans-IL-15 expression on iDCs, Ad.DCs, and mDCs was examined with confocal microscopy. (Top panels) Optical cross-sections. (Bottom panels) Reconstructed (3-dimensional) images of 10 0.4-μm-thick optical sections. (A-B) Results are representative of 3 experiments. Lysates of 24 hours (C-D) and supernatants of 24 hours and 48 hours (E-F) cultured iDCs, Ad.DCs, and mDCs were tested for cell-associated (plasma membrane-bound) and soluble TNF and IL-15 by ELISA and cytokine multiplex assays, respectively. (B,D) TNF data are mean ± SEM (pg/mL) of 7 independent experiments. (C,E) IL-15 data are mean ± SEM (pg/mL) of 3 independent experiments. *P < .05.